Triple Amplification Strategy for the Improved Efficiency of a Microplate-Based Assay for the Chemiluminescent Detection of DNAстатья

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Дата последнего поиска статьи во внешних источниках: 29 мая 2019 г.

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[1] Kolosova A. Y., Sakharov I. Y. Triple amplification strategy for the improved efficiency of a microplate-based assay for the chemiluminescent detection of dna // Analytical Letters. — 2019. — Vol. 52, no. 8. — P. 1352–1362. Nowadays, considerable research efforts are focused on advancing DNA detection methodology. Various nanoparticles (NPs), which are currently the most widely employed solid-phase carriers for nucleic acid assays, have a number of essential drawbacks. Microtiter plates provide a simple and economical alternative to the NPs. The present paper reports the development of sandwich assay for DNA detection using microtiter plate as a solid carrier. Capture oligonucleotide modified with fluorescein was bound to the antifluorescein antibody adsorbed on the polystyrene microplate surface. Hepatitis B virus (HBV) DNA fragment was used as a model analyte. To improve the assay sensitivity, the biotinylated reporter oligonucleotide and streptavidin-horseradish polyperoxidase (polyHRP) conjugate were used as an amplified detection system. Additional amplification was achieved due to the fact that peroxidase activity was measured by chemiluminescent method using 3-(10’-phenothiazinyl)propane-1-sulfonate/N-morpholinopyridine pair as a enhancer. Detection limit of the developed assay was 0.9 pM, a linear range – from 0.9 to 100 pM. This method can be used as a platform for the development of sensitive bio(DNA/apta)assays. [ DOI ]

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